A multiplex real-time PCR method based on fluorescent TaqMan® probes was developed for the simultaneous detection of the tomato pathogenic bacteria Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato and bacterial spot-causing xanthomonads. The specificity of the multiplex assay was validated on 44 bacterial strains, including 32 target pathogen strains as well as closely related species and nontarget tomato pathogenic bacteria. The designed multiplex real-time PCR showed high sensitivity when positive amplification was observed for one pg of bacterial DNA in the cases of Clavibacter michiganensis subsp. michiganensis and Pseudomonas syringae pv. tomato bacteria and 100 pg for bacterial spot-causing xanthomonads. The reliability of the developed multiplex real-time PCR assay for in planta detection was verified by recognition of the target pathogens in 18 tomato plants artificially inoculated by each of the target bacteria and tomato samples from production greenhouses.
- MeSH
- Actinobacteria genetika izolace a purifikace fyziologie MeSH
- kvantitativní polymerázová řetězová reakce * MeSH
- prostředí kontrolované MeSH
- Pseudomonas syringae genetika izolace a purifikace fyziologie MeSH
- Solanum lycopersicum růst a vývoj mikrobiologie MeSH
- Xanthomonas genetika izolace a purifikace fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
In this report we describe a specific transfer of carbenicillin and cephaloridine resistance determinants from two different strains of Stenotrophomonas maltophilia: No. 215 and 221 isolated from two critically ill patients treated in different Intensive Care Units of a large University Hospital in Ostrava, Czech Republic. These strains were resistant to flouroquinolones and the following beta-lactam drugs: carbenicillin, cephaloridine, cefotaxime, ceftazidime, cefepime, imipenem, meropenem and aztreonam. Both strains transferred carbenicillin and cephaloridine resistance determinants, with rather different frequency, to Proteus mirabilis P-38. All carbenicillin-selected transconjugants were found by an indirect selection method to be co-resistant to cephaloridine only. In a second cycle of transfers Proteus mirabilis R+ strains directly transferred carbenicillin and cephalothin determinants to Escherichia coli K-12 No. 185 nal+ lac+ recipient strain.
- MeSH
- antibakteriální látky farmakologie MeSH
- antiinfekční látky farmakologie MeSH
- beta-laktamasy analýza metabolismus MeSH
- beta-laktamová rezistence * MeSH
- cefaloridin farmakologie MeSH
- fluorochinolony MeSH
- jednotky intenzivní péče MeSH
- karbenicilin farmakologie MeSH
- kultivační média MeSH
- lidé MeSH
- mnohočetná léková rezistence * MeSH
- Proteus mirabilis účinky léků genetika MeSH
- Xanthomonas účinky léků genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
